Escherichia coli|194754 |BNCC

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Enterococcus hirae

Subculture procedure	① Prepare two of the aforementioned plates (pre-equilibrate in an anaerobic environment for 24 hours). ② Disinfect the surface of the ampoule vial, open it within a biosafety cabinet, flame the top using an alcohol lamp, then promptly drop sterile water onto it to induce cracking, followed by breaking it with forceps. ③ Aspirate 0.5 mL of sterile water (pre-equilibrated in an anaerobic environment for 24 hours), inject it into the lyophilized vial, thoroughly resuspend the bacterial powder, then inoculate the suspension onto two plates (200 µL per Agar plate) and spread evenly. ④ Incubate the plates under the specified culture conditions mentioned above. The strain is ready for use once growth is observed.
Growth conditions	37 ℃; 24-48 hours; anaerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	The colony diameter is 1-2mm, circular, with neat edges, opaque, milky white on the front, raised in the middle, smooth and bright on the surface, moist texture, easy to pick up, G+(blue purple), cocci, purity: pure
Sharing mode	Public welfare sharing

1.Description

1. Name：Enterococcus hirae

2. BNCC No.：194754

3. Biosafety level：4

2.Storage conditions：

Storage of freezed dried ampoule and agar slant at 2°C to 8°C

3.Growth Conditions

1. MRS medium + 0.05% cysteine:(MRS medium: cheese peptone 10.0g, beef powder 8.0g, yeast powder 4.0g, glucose 20.0g, magnesium sulfate 0.2g, sodium acetate 5.0g, triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.05g, Tween 80 1.0g, distilled water 1000mL. pH6.2±0.2.121 ℃,15min.)

2. Atmosphere： anaerobic

3. Temperature：37 ℃

4.Notes：

1.Normal culturing time, 24-48hours for bacterial, 72 hours for yeast, 5-7days for mould, 7-10days for fungal.

2.Agar slant shall be inoculated asap, and do not keep the storage for more than 3 months.

3.Please recover the strains in strict accordance with this instruction, otherwise the replacement of the strain are not be available in case of viability loss caused by different media or growth conditions.

4.Waste generated from the handling process should be discarded after high-pressure sterilization

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